Integrating Energy Calculations with Functional Assays to Decipher the Specificity of G Protein-RGS Protein Interactions

Overview

Journal Nat Struct Mol Biol

Specialties Cell Biology
Molecular Biology

Date 2011 Jun 21

PMID 21685921

Citations 12

Authors

Mickey Kosloff

Amanda M Travis

Dustin E Bosch

David P Siderovski

Vadim Y Arshavsky

Affiliations

Soon will be listed here.

Abstract

The diverse Regulator of G protein Signaling (RGS) family sets the timing of G protein signaling. To understand how the structure of RGS proteins determines their common ability to inactivate G proteins and their selective G protein recognition, we combined structure-based energy calculations with biochemical measurements of RGS activity. We found a previously unidentified group of variable 'Modulatory' residues that reside at the periphery of the RGS domain-G protein interface and fine-tune G protein recognition. Mutations of Modulatory residues in high-activity RGS proteins impaired RGS function, whereas redesign of low-activity RGS proteins in critical Modulatory positions yielded complete gain of function. Therefore, RGS proteins combine a conserved core interface with peripheral Modulatory residues to selectively optimize G protein recognition and inactivation. Finally, we show that our approach can be extended to analyze interaction specificity across other large protein families.

Citing Articles

Bluues_cplx: Electrostatics at Protein-Protein and Protein-Ligand Interfaces.

Soler M, Yakout R, Ozkilinc O, Esposito G, Rocchia W, Klein C Molecules. 2025; 30(1.

PMID: 39795215 PMC: 11722155. DOI: 10.3390/molecules30010159.

Evolution of homo-oligomerization of methionine S-adenosyltransferases is replete with structure-function constrains.

Kleiner D, Shapiro Tuchman Z, Shmulevich F, Shahar A, Zarivach R, Kosloff M Protein Sci. 2022; 31(7):e4352.

PMID: 35762725 PMC: 9202080. DOI: 10.1002/pro.4352.

Residue-level determinants of RGS R4 subfamily GAP activity and specificity towards the G subfamily.

Asli A, Higazy-Mreih S, Avital-Shacham M, Kosloff M Cell Mol Life Sci. 2021; 78(17-18):6305-6318.

PMID: 34292354 PMC: 11072900. DOI: 10.1007/s00018-021-03898-4.

Structural design principles for specific ultra-high affinity interactions between colicins/pyocins and immunity proteins.

Shushan A, Kosloff M Sci Rep. 2021; 11(1):3789.

PMID: 33589691 PMC: 7884437. DOI: 10.1038/s41598-021-83265-2.

Allosteric Pathways Originating at Cysteine Residues in Regulators of G-Protein Signaling Proteins.

Liu Y, Vashisth H Biophys J. 2020; 120(3):517-526.

PMID: 33347886 PMC: 7895990. DOI: 10.1016/j.bpj.2020.12.010.

References

Kortemme T, Joachimiak L, Bullock A, Schuler A, Stoddard B, Baker D . Computational redesign of protein-protein interaction specificity. Nat Struct Mol Biol. 2004; 11(4):371-9. DOI: 10.1038/nsmb749. View

Li W, Keeble A, Giffard C, James R, Moore G, Kleanthous C . Highly discriminating protein-protein interaction specificities in the context of a conserved binding energy hotspot. J Mol Biol. 2004; 337(3):743-59. DOI: 10.1016/j.jmb.2004.02.005. View

Huang H, Yuan H . The conserved asparagine in the HNH motif serves an important structural role in metal finger endonucleases. J Mol Biol. 2007; 368(3):812-21. DOI: 10.1016/j.jmb.2007.02.044. View

Doudeva L, Huang H, Hsia K, Shi Z, Li C, Shen Y . Crystal structural analysis and metal-dependent stability and activity studies of the ColE7 endonuclease domain in complex with DNA/Zn2+ or inhibitor/Ni2+. Protein Sci. 2006; 15(2):269-80. PMC: 2242460. DOI: 10.1110/ps.051903406. View

Wang Q, Liu M, Mullah B, Siderovski D, Neubig R . Receptor-selective effects of endogenous RGS3 and RGS5 to regulate mitogen-activated protein kinase activation in rat vascular smooth muscle cells. J Biol Chem. 2002; 277(28):24949-58. DOI: 10.1074/jbc.M203802200. View

Siderovski D, Hessel A, Chung S, Mak T, Tyers M . A new family of regulators of G-protein-coupled receptors?. Curr Biol. 1996; 6(2):211-2. DOI: 10.1016/s0960-9822(02)00454-2. View

Slep K, Kercher M, Wieland T, Chen C, Simon M, Sigler P . Molecular architecture of Galphao and the structural basis for RGS16-mediated deactivation. Proc Natl Acad Sci U S A. 2008; 105(17):6243-8. PMC: 2359805. DOI: 10.1073/pnas.0801569105. View

Ross E, Wilkie T . GTPase-activating proteins for heterotrimeric G proteins: regulators of G protein signaling (RGS) and RGS-like proteins. Annu Rev Biochem. 2000; 69:795-827. DOI: 10.1146/annurev.biochem.69.1.795. View

Watson N, Linder M, Druey K, Kehrl J, Blumer K . RGS family members: GTPase-activating proteins for heterotrimeric G-protein alpha-subunits. Nature. 1996; 383(6596):172-5. DOI: 10.1038/383172a0. View

10.

Wieland T, Bahtijari N, Zhou X, Kleuss C, Simon M . Polarity exchange at the interface of regulators of G protein signaling with G protein alpha-subunits. J Biol Chem. 2000; 275(37):28500-6. DOI: 10.1074/jbc.M004187200. View

11.

Ko T, Liao C, Ku W, Chak K, Yuan H . The crystal structure of the DNase domain of colicin E7 in complex with its inhibitor Im7 protein. Structure. 1999; 7(1):91-102. DOI: 10.1016/s0969-2126(99)80012-4. View

12.

Ingi T, Krumins A, Chidiac P, Brothers G, Chung S, Snow B . Dynamic regulation of RGS2 suggests a novel mechanism in G-protein signaling and neuronal plasticity. J Neurosci. 1998; 18(18):7178-88. PMC: 6793237. View

13.

Kimple A, Soundararajan M, Hutsell S, Roos A, Urban D, Setola V . Structural determinants of G-protein alpha subunit selectivity by regulator of G-protein signaling 2 (RGS2). J Biol Chem. 2009; 284(29):19402-11. PMC: 2740565. DOI: 10.1074/jbc.M109.024711. View

14.

Laroche G, Giguere P, Roth B, Trejo J, Siderovski D . RNA interference screen for RGS protein specificity at muscarinic and protease-activated receptors reveals bidirectional modulation of signaling. Am J Physiol Cell Physiol. 2010; 299(3):C654-64. PMC: 2944319. DOI: 10.1152/ajpcell.00441.2009. View

15.

Sowa M, He W, Wensel T, Lichtarge O . A regulator of G protein signaling interaction surface linked to effector specificity. Proc Natl Acad Sci U S A. 2000; 97(4):1483-8. PMC: 26460. DOI: 10.1073/pnas.030409597. View

16.

Kuhlmann U, Pommer A, MOORE G, James R, Kleanthous C . Specificity in protein-protein interactions: the structural basis for dual recognition in endonuclease colicin-immunity protein complexes. J Mol Biol. 2000; 301(5):1163-78. DOI: 10.1006/jmbi.2000.3945. View

17.

Koelle M, Horvitz H . EGL-10 regulates G protein signaling in the C. elegans nervous system and shares a conserved domain with many mammalian proteins. Cell. 1996; 84(1):115-25. DOI: 10.1016/s0092-8674(00)80998-8. View

18.

Schreiber G, Keating A . Protein binding specificity versus promiscuity. Curr Opin Struct Biol. 2010; 21(1):50-61. PMC: 3053118. DOI: 10.1016/j.sbi.2010.10.002. View

19.

Hollinger S, Hepler J . Cellular regulation of RGS proteins: modulators and integrators of G protein signaling. Pharmacol Rev. 2002; 54(3):527-59. DOI: 10.1124/pr.54.3.527. View

20.

Berman D, Wilkie T, GILMAN A . GAIP and RGS4 are GTPase-activating proteins for the Gi subfamily of G protein alpha subunits. Cell. 1996; 86(3):445-52. DOI: 10.1016/s0092-8674(00)80117-8. View